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Production of a cytotoxic cationic antibacterial peptide in Escherichia coli using SUMO fusion partner

Abstract

Antibacterial peptide CM4 (ABP-CM4) is a small cationic peptide with broad-spectrum activities against bacteria, fungi, and tumor cells, which may possibly be used as an antimicrobial agent. We report here the application of small ubiquitin-related modifier (SUMO) fusion technology to the expression and purification of cationic antibacterial peptide ABP-CM4. The fusion protein expressed in a soluble form was purified to a purity of 90% by Ni-IDA chromatography and 112 mg protein of interest was obtained per liter of fermentation culture. After the SUMO–CM4 fusion protein was cleaved by the SUMO protease at 30 °C for 1 h, the cleaved sample was re-applied to a Ni-IDA. Finally, about 24 mg recombinant CM4 was obtained from 1 l fermentation culture with no less than 96% purity and the recombinant CM4 had similar antimicrobial properties to the synthetic CM4. Thus, the SUMO-mediated peptide expression and purification system potentially could be employed for the production of recombinant cytotoxic peptides.

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Acknowledgements

This work was supported by the Grants of Nanjing Normal University and Jiangsu Province Graduate Innovation Project (No. CX07S-020z) administered by Prof. Zhang.

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Correspondence to Shuang Quan Zhang.

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Li, J.F., Zhang, J., Song, R. et al. Production of a cytotoxic cationic antibacterial peptide in Escherichia coli using SUMO fusion partner. Appl Microbiol Biotechnol 84, 383–388 (2009). https://doi.org/10.1007/s00253-009-2109-2

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Keywords

  • Antibacterial peptide
  • ABP-CM4
  • SUMO
  • Escherichia coli